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M. C. Giovingo, R. D. McCarty, R. M. Beverley, M. J. Nolan, J. R. Samples, B. Y. J. T. Yue, P. A. Knepper; Soluble CD44 Increases Outflow Resistance in Porcine Organ Cultures. Invest. Ophthalmol. Vis. Sci. 2010;51(13):5838.
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CD44 plays major roles in multiple physiological processes.The soluble CD44 (sCD44) concentration is significantly increasedin the aqueous humor of primary open-angle glaucoma (POAG).Increased sCD44 in mouse eyes causes an increase in intraocularpressure (IOP). The purpose of this study was to determine ifsCD44 changes outflow resistance in porcine anterior segmentorgan culture and profiles of lipid raft proteins of the trabecularmeshwork (TM).
sCD44 was purified from human serum using anion exchange, hyaluronicacid (HA) affinity chromatography and immunoprecipitation. Anteriorsegments of porcine eyes were placed in organ culture and perfusedwith Dulbecco’s modified eagle medium (DMEM). Flow rateswere measured in eyes treated with sCD44, HA or DMEM. Perturbationof lipid raft containing proteins was assessed by Optiprep densitygradient and Western blot analysis of dissected TM.
Flow rates are expressed as percent change (+/-) from the baseline.Rates significantly decreased from baseline in HA and sCD44infused eyes.
Analysis of lipid raft containing proteinsof sCD44 infused porcine microdissected TMs and primary culturesof TM cells revealed a decrease in annexin 2 and an increasein caveolin-1 compared to that of controls.
Infusion of sCD44 significantly decreased outflow facility inporcine eyes. Notably, sCD44 caused a decrease in annexin 2and an increase in caveolin-1 which may influence outflow resistanceand the regulation of IOP.
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